Dermacentor Nitens Research Articles

Morphological and molecular data suggest the presence of cryptic diversity within Dermacentor nitens (Acari: Ixodidae) in Colombia</p >

Of the 44 species within the genus Dermacentor (Acari: Ixodidae) worldwide, nine are found in the Neotropics and four are endemic to this region. In Colombia, located in the northwestern corner of South America, there are two species of the genus: Dermacentor imitans restricted to the trans-Andean Pacific region, and Dermacentor nitens widely distributed in the country. Elsewhere, D. nitens is found in both the Nearctic and Neotropics, from southern USA to northern Argentina. In the Neotropics, D. nitens is widely distributed including records in Caribbean islands. Molecular analyses of D. nitens collected in the Amazon region of Colombia have shown genetic differences with specimens from other localities of the country, although morphological differences have not been explored. We performed morphological and molecular comparisons of individuals from two natural regions of Colombia: the Orinoquia, and the Amazon. The morphological and morphometric results show that females and males are grouped in two morphotypes: morphotype I from the Orinoquia region, and morphotype II from Leticia in the Amazon region. The molecular analyses supported the morphological results and showed the formation of two well-differentiated sister clades within D. nitens, with divergences between both morphotypes of 3–4% and 5–6% for the 16S rRNA and COI genes, respectively. Species delimitation analyzes and the Poisson Tree Processes model tentatively suggest the presence of two species within the sequences associated with D. nitens in Colombia. The first represented by the sequences from the Orinoquia region evaluated here, together with sequences from other regions of Colombia and other Neotropical countries such as Brazil, Panama, and French Guiana. The second is represented by the sequences of individuals from Leticia, in the Amazon region of Colombia. The results highlight that D. nitens might represent a species complex, but further sampling, experimental crosses, and studies at the level of complete genomes should be performed to obtain additional evidence for species recognition.

Hard ticks (Ixodida: Ixodidae) in the Colombian Caribbean harbor the Jingmen tick virus: an emerging arbovirus of public health concern

BackgroundTicks are obligate hematophagous ectoparasites involved in transmitting viruses of public health importance. The objective of this work was to identify the Jingmen tick virus in hard ticks from the Colombian Caribbean, an arbovirus of importance for public health.MethodsTicks were collected in rural areas of Córdoba and Cesar, Colombia. Taxonomic identification of ticks was carried out, and pools of 13 individuals were formed. RNA extraction was performed. Library preparation was performed with the MGIEasy kit, and next-generation sequencing (NGS) with MGI equipment. Bioinformatic analyses and taxonomic assignments were performed using the Galaxy platform, and phylogenetic analyses were done using IQ-TREE2.ResultsA total of 766 ticks were collected, of which 87.33% (669/766) were Rhipicephalus microplus, 5.4% (42/766) Dermacentor nitens, 4.2% (32/766) Rhipicephalus linnaei, and 3.0% (23/766) Amblyomma dissimile. Complete and partial segments 1, 2, 3, and 4 of Jingmen tick virus (JMTV) were detected in the metatranscriptome of the species R. microplus, D. nitens, and A. dissimile. The JMTVs detected are phylogenetically related to JMTVs detected in Aedes albopictus in France, JMTVs detected in R. microplus in Trinidad and Tobago, JMTVs in R. microplus and A. variegatum in the French Antilles, and JMTVs detected in R. microplus in Colombia. Interestingly, our sequences clustered closely with JMTV detected in humans from Kosovo.ConclusionsJMTV was detected in R. microplus, D. nitens, and A. dissimile. JMTV could pose a risk to humans. Therefore, it is vital to establish epidemiological surveillance measures to better understand the possible role of JMTV in tropical diseases.Graphical

Diversity of midgut microbiota in ticks collected from white-tailed deer (Odocoileus virginianus) from northern Mexico.

Ticks host different pathogens as endosymbiont and nonpathogenic microorganisms and play an important role in reproductive fitness and nutrient provision. However, the bacterial microbiomes of white-tailed deer ticks have received minimal attention. This study aimed to examine the bacterial microbiome of ticks collected from Odocoileus virginianus on the Mexico-United States border to assess differences in microbiome diversity in ticks of different species, sexes, and localities. Five different tick species were collected: Rhipicephalus microplus, Dermacentor nitens, Otobius megnini, Amblyomma cajennense, and A. maculatum. The tick microbiomes were analyzed using next-generation sequencing. Among all tick species, the most predominant phylum was Proteobacteria, followed by Actinobacteria and Firmicutes. The ticks from Tamaulipas and Nuevo León presented the highest bacterial species diversity. Acinetobacter johnsonii and A. lwoffii were the common bacterial species in the microbiome of all ticks, Coxiella were present in R. microplus, and Dermacentor nitens also exhibited a Francisella-like endosymbiont. The microbiome of most females in D. nitens was less diverse than that of males, whereas R. microplus occurs in females, suggesting that microbiome diversity is influenced by sex. In the bacterial communities of A. maculatum and O. megnini, Candidatus Midichloria massiliensis, and Candidatus Endoecteinascidia fumentensis were the most predominant endosymbionts. These results constitute the initial report on these bacteria, and this is also the first study to characterize the microbiome of O. megnini.

Efficacy of the Vaccine Candidate Based on the P0 Peptide against Dermacentor nitens and Ixodes ricinus Ticks.

The control of ticks through vaccination offers a sustainable alternative to the use of chemicals that cause contamination and the selection of resistant tick strains. However, only a limited number of anti-tick vaccines have reached commercial realization. In this sense, an antigen effective against different tick species is a desirable target for developing such vaccines. A peptide derived from the tick P0 protein (pP0) conjugated to a carrier protein has been demonstrated to be effective against the Rhipicephalus microplus, Rhipicephalus sanguineus, and Amblyomma mixtum tick species. The aim of this work was to assess the efficacy of this peptide when conjugated to the Bm86 protein against Dermacentor nitens and Ixodes ricinus ticks. An RNAi experiment using P0 dsRNA from I. ricinus showed a dramatic reduction in the feeding of injected female ticks on guinea pigs. In the follow-up vaccination experiments, rabbits were immunized with the pP0-Bm86 conjugate and challenged simultaneously with larvae, nymphs, and the adults of I. ricinus ticks. In the same way, horses were immunized with the pP0-Bm86 conjugate and challenged with D. nitens larva. The pP0-Bm86 conjugate showed efficacies of 63% and 55% against I. ricinus and D. nitens ticks, respectively. These results, combined with previous reports of efficacy for this conjugate, show the promising potential for its development as a broad-spectrum anti-tick vaccine.

First report of Amblyomma sculptum (Amblyomma cajennense complex) in a Brazilian state classified as a silent area for human rickettsiosis.

Studies on ticks of public health concern in equine husbandry are scarce in the Northeastern region of Brazil. This study aimed to investigate the presence of ticks on horses in the State of Alagoas, which is classified as a silent area for human rickettsiosis. Ticks infesting horses were collected using anatomical tweezers or a commercial hook and kept in ethanol-labeled tubes for taxonomic identification. A total of 2,238 ticks were found. Ticks were identified as 2,215 (98.89%, 95% CI: 98.41-99.28) Dermacentor nitens, 19 (0.98%, 95% CI: 0.05-1.38) Amblyomma sculptum, and 4 (0.18%; 95% CI: 0.007-0.46) Rhipicephalus microplus. This is the first study to report A. sculptum and D. nitens in the State of Alagoas. The presence of A. sculptum should draw the attention of public health managers once Alagoas State is considered a silent area for rickettsial diseases, which means the absence of local surveillance programs for these pathogens.

Diversity of the Bacterial and Viral Communities in the Tropical Horse Tick, Dermacentor nitens, in Colombia.

Ticks are obligatory hematophagous ectoparasites that transmit pathogens among various vertebrates, including humans. The microbial and viral communities of ticks, including pathogenic microorganisms, are known to be highly diverse. However, the factors driving this diversity are not well understood. The tropical horse tick, Dermacentor nitens, is distributed throughout the Americas and it is recognized as a natural vector of Babesia caballi and Theileria equi, the causal agents of equine piroplasmosis. In this study, we characterized the bacterial and viral communities associated with partially fed Dermacentor nitens females collected using a passive survey on horses from field sites representing three distinct geographical areas in the country of Colombia (Bolivar, Antioquia, and Cordoba). RNA-seq and sequencing of the V3 and V4 hypervariable regions of the 16S rRNA gene were performed using the Illumina-Miseq platform (Illumina, San Diego, CA, USA). A total of 356 operational taxonomic units (OTUs) were identified, in which the presumed endosymbiont, Francisellaceae/Francisella spp., was predominantly found. Nine contigs corresponding to six different viruses were identified in three viral families: Chuviridae, Rhabdoviridae, and Flaviviridae. Differences in the relative abundance of the microbial composition among the geographical regions were found to be independent of the presence of Francisella-like endosymbiont (FLE). The most prevalent bacteria found in each region were Corynebacterium in Bolivar, Staphylococcus in Antioquia, and Pseudomonas in Cordoba. Rickettsia-like endosymbionts, mainly recognized as the etiological agent of rickettsioses in Colombia, were detected in the Cordoba samples. Metatranscriptomics revealed 13 contigs containing FLE genes, suggesting a trend of regional differences. These findings suggest regional distinctions among the ticks and their bacterial compositions.

Clinical and epidemiological aspects of the infection by Babesia, Theileria and Trypanosoma species in horses from northeastern Colombia

Piroplasmosis and trypanosomiasis are debilitating diseases of great economic impact on the equine industry of Latin America. Considering the lack of studies in the northeastern part of Colombia, this study aimed to determine the epidemiological, clinical and genetic features associated with infection of the Babesia, Theileria, and Trypanosoma species in horses from this geographical area. Two hundred and eighty horses from the Arauca, Meta, and Santander departments were molecularly analyzed for infection with Babesia caballi, Theileria equi, Trypanosoma evansi, and Trypanosoma vivax. Furthermore, clinical, epidemiological and entomological analyses were performed on the data sets. Molecular analysis showed 25.7% and 3.9% prevalence for T. equi and T. evansi, respectively, without positive animals for B. caballi and T. vivax. There were no differences in the prevalence of T. equi between departments, whereas T. evansi was detected exclusively in Santander. A total of 633 ticks were collected from 72 horses across the three departments, with 84.7% corresponding to Dermacentor nitens, 10.9% to Amblyomma cajennense (sensu lato) (s.l). and 4.4% to Rhipicephalus microplus. For T. equi, genetic analyses showed that Colombian isolates belong to genotype C of species, along with sequences of Brazil and Mexico. Epidemiological analysis revealed a significant association between tick infestation and lack of vector control with molecular infection of T. equi, whereas clinical analysis revealed a significant reduction in packed cell volume, red blood cells, and mean corpuscular volume in positive animals to this pathogen. Furthermore, molecular infection by T. evansi was associated with epidemiological characteristics in the Santander department. In conclusion, our analysis revealed a moderate infection rate by T. equi of genotype C in horses from northeastern Colombia, which affects their clinical conditions. Control of ticks and treatment of symptomatic animals should be considered to reduce the economic impact associated with these infections in the equine industry.

Circulating oxidative stress and acute phase protein levels in horses infested with ticks.

Ticks have saliva rich in immunoregulatory molecules that interfere with the host's physiology in order to feed. This study aimed to evaluate the concentration of acute phase proteins and circulating oxidative stress in response to infestation by Amblyomma sculptum and Dermacentor nitens in two breed horses, Mangalarga Marchador and Breton Postier, to define resistance or susceptibility to ticks. Among the oxidative stress markers, we observed lower malondialdehyde and nitric oxide in horses with tick infestation, consequently not altering the antioxidant enzymes. Breton Postier with tick infestation showed a reduction in the ferric reducing ability of plasma (FRAP), which may be due to lower feeding of the host due to the stress caused by the infestation or even to sequestration of components induced by the tick during blood feeding. The alpha-1-antitrypsin, an acute phase protein, showed an increase in Mangalarga Marchador with tick infestation; curiously it is related to a protective action against tissue damage, pathogens and parasites. We could assume that Mangalarga Marchador showed a better response to ticks when compared to Breton Postier. However, it is still early to define the resistance or susceptibility to ticks, as we did not observe significant changes in most of the analyzed variables. Further studies are needed to understand the compounds and mechanisms of action of the tick saliva in the acute phase proteins and the possible relationships of oxidative stress in the host and the tick during blood feeding.

Detection of Rickettsia spp. in Animals and Ticks in Midwestern Brazil, Where Human Cases of Rickettsiosis Were Reported.

Brazilian spotted fever (BSF) is the most important tick-borne diseases affecting humans in Brazil. Cases of BSF have recently been reported in the Goiás state, midwestern Brazil. All cases have been confirmed by reference laboratories by seroconversion to Rickettsia rickettsii antigens. Because serological cross-reactions among different rickettsial species that belong to the spotted fever group (SFG) are common, the agent responsible for BSF cases in Goiás remains unknown. From March 2020 to April 2022, ticks and plasma were collected from dogs, horses and capybaras (Hydrochoerus hydrochaeris), and from the vegetation in an area where BSF cases have been reported and two areas under epidemiological surveillance in Goiás. Horses were infested by Amblyomma sculptum, Dermacentor nitens and Rhipicephalus microplus; dogs by Rhipicephalus sanguineus sensu lato (s.l.), Amblyomma ovale and A. sculptum, and capybaras by A. sculptum and Amblyomma dubitatum. Adults of A. sculptum, A. dubitatum, Amblyomma rotundatum and immature stages of A. sculptum and A. dubitatum, and Amblyomma spp. were collected from the vegetation. DNA of Rickettsia that did not belong to the SFG was detected in A. dubitatum, which was identified by DNA sequencing as Rickettsia bellii. Seroreactivity to SFG and Rickettsia bellii antigens was detected in 25.4% (42/165) of dogs, 22.7% (10/44) of horses and 41.2% (7/17) of capybaras, with higher titers for R. bellii in dogs and capybaras. The seropositivity of animals to SFG Rickettsia spp. antigens demonstrates the circulation of SFG rickettsiae in the region. Further research is needed to fully determine the agent responsible for rickettsiosis cases in this area.

Analysis of the tick communities associated to domestic mammals in rural areas of the Yungas montane forest from Argentina.

The aim of this work was to describe the tick community associated to domestic mammals in rural areas from the Yungas lower montane forest of Argentina. The circulation of tick-borne pathogens was also analyzed. Samples of ticks parasitizing cattle, horses, sheep and dogs were carried out in different seasons, and questing ticks were collected from vegetation and analyzed to detect the presence of Rickettsia, Ehrlichia, Borrelia and Babesia by a battery of different PCRs. The structure of the tick communities was analyzed through the Chao1 species richness estimator, the Shannon-Wiener index and the Horn index of community similarity. Eight tick species were collected in the study area: Amblyomma sculptum, Rhipicephalus microplus, Amblyomma hadanii, Dermacentor nitens, Amblyomma ovale, Haemaphysalis juxtakochi, Ixodes pararicinus and Rhipicephalus sanguineus sensu stricto. However, A. sculptum was by far the dominant species in the tick assemblages analyzed, and this was reflected in the low diversity values obtained. Dermacentor nitens, A. sculptum and R. microplus were the three species associated to horses. The predominance of A. sculptum was also observed in the tick samples obtained from dogs, even on two tick species, namely A. ovale and R. sanguineus s.s., which have dogs as the principal domestic host. Rhipicephalus microplus and A. sculptum were the most abundant ticks on cattle, while few specimens of I. pararicinus, A. hadanii and D. nitens were found on bovines. Dermacentor nitens ticks were found to be infected with B. caballi, which indicate the circulation of this pathogen of horses in the Yungas area. The detection of a strain of Borrelia sp. belonging to the B. burgdorferi s.l. complex in I. pararicinus is consistent with previous findings made in Argentina, but the public health relevance of this vector-microorganism association is far from being similar to that occurs in the northern hemisphere because there are practically no records of these tick species parasitizing humans in South America. The tick community of rural areas of the Yungas lower montane forest is composed by species which are potential vectors of pathogenic microorganism with veterinary and public health importance, circulating in a human-wildlife-livestock interface.

Comparison of Seroprevalence and Identification of Risk Factors for Theileria equi in Horses From Vector-Free and Infested Areas in Southern Brazil

The apicomplexan hemoprotozoan Theileria equi is a tick-borne pathogen that causes disease in equids, and together with Babesia caballi, causes equine piroplasmosis (EP). Many ticks are associated with EP around the world, and in South America three species may be related: Dermacentor nitens, Amblyomma sculptum, and Rhipicephalus microplus, as they are commonly found in horses. Among the species cited above, only R. microplus is found in Rio Grande do Sul state, Southern Brazil. In addition, this state has the only area legally free of R. microplus in Brazil. This study aimed to compare the seroprevalence for T. equi on farms located in a tick-free area (23 farms, 215 horses) and a tick-infested (25 farms, 141 horses) adjacent areas, as well as to identify potential risk factors for exposure to T. equi. Blood was collected from 356 horses from both areas, and later an indirect enzyme-linked immunosorbent assay was performed to detect anti-T. equi antibodies. Besides the blood collection, questionnaires and interviews were conducted in an attempt to identify potential risk factors. The R. microplus-free zone had 6% seropositive horses against 70% in the tick-infested zone. Previous observation of ticks on horses of the herd increased four times the odds of a horse to be seropositive for T. equi, and by three times if ticks were observed on cattle which share paddocks with horses on the farm. The results showed a large difference in T. equi seroprevalence between tick-infested and free areas, and also emphasized the relevance of R. microplus in the epidemiology of T. equi. The study also reveals the potential of the establishment of a T. equi-free zone for horse breeding in Southern Brazil.

Parasitismo em local incomum de Dermacentor nitens (Acari: Ixodidae) em um cavalo

ABSTRACT: Dermacentor nitens Neumann is the most common tick species infesting horses in the main Brazilian biomes. It has a predilection to attach to horse the ears, the nasal diverticulum, perineal and perianal regions. The infestations can generate severe damage in the ears, anemia, and the tick also acts as vector of Babesia caballi (Nuttall and Strickland), the causative agent of equine babesiosis. Our study describes unusual parasitism site of D. nitens on a female cross breed horse, approximately ten years old that presented high parasitism by ticks on the perineal and perianal region, ears, and the left eye orbit region, where an enucleation process had been performed a few years earlier. To our knowledge this is the first report of D. nitens parasitism on a formerly enucleated eye orbit.

Molecular detection of tick-borne rickettsial pathogens in ticks collected from domestic animals from Cauca, Colombia

Some hard ticks’ species can act as vectors of a wide variety of pathogens of human and animal importance such as Anaplasma, Ehrlichia and Rickettsia spp. In Colombia, a total of forty-six tick species have been described, and some of them have been implicated as vectors of some infectious agents. The department of Cauca is one of the thirty-two departments of Colombia. Most of its population lives in rural areas and depends on agriculture as the main economic activity, favoring exposure to ticks and tick-borne pathogens. Thus, the present study aimed to determine the tick species and tick-borne pathogens circulating in this region. From August to November 2017, ticks were collected from dogs, horses and cattle from eight rural areas of four municipalities in the department of Cauca. All collected ticks were classified according to taxonomic keys and organized in pools. DNA was extracted from all tick pools for molecular confirmation of tick species and detection of Anaplasma, Ehrlichia and Rickettsia spp. A total of 2809 ticks were collected which were grouped in 602 pools. Ticks were morphologically identified as Amblyomma cajennense sensu lato, Dermacentor nitens, Rhipicephalus microplus and Rhipicephalus sanguineus sensu lato. The molecular identity of A. cajennense s.l. was confirmed as Amblyomma patinoi. A total of 95% of the pools scored positive for members of the Anaplasmataceae family, of which, 7.8% and 7.3% were positive to Anaplasma and Ehrlichia spp., respectively, being identified as Anaplasma marginale, Ehrlichia minasensis and Ehrlichia canis; and 16.1% were positive for Rickettsia spp. with high identity for Rickettsia asembonensis, Rickettsia felis and Candidatus Rickettsia senegalensis. This is the first report describing the natural infection of ticks with rickettsial pathogens and the occurrence of A. patinoi ticks in Cauca department, Colombia.

Cuticular Lipids as a First Barrier Defending Ixodid Ticks against Fungal Infection.

The chemical composition of tick cuticles acts as a barrier to pathogens and may limit infection by entomopathogenic fungi. This study characterized the cuticular neutral lipids (NL) and hydrocarbons (HCs) of four ixodid ticks that are widely distributed in Brazil. HC extracts were analyzed by gas chromatography-mass spectrometry and used to challenge Beauveria bassiana IP361 and Metarhizium robertsii IP146; the effect of cuticular extracts in fungal growth were evaluated by disk diffusion and conidial viability assays. In addition, conidial germination on the tick cuticle was evaluated by scanning electron microscopy, and NL from ticks treated with fungi were assessed by thin layer chromatography. Six HCs were exclusively identified in Amblyomma sculptum. Additionally, cuticle extracts from Dermacentor nitens and A. sculptum inhibited the growth of M. robertsii IP146 and reduced conidial germination of B. bassiana IP361 to 70% and 49%, respectively; the same extracts also produced cytotoxic effects, with conidial death above 30% and 60%. Electron micrographs showed a delayed germination of conidia incubated for 48 h or 72 h on D. nitens and A. sculptum. The lipid profile of A. sculptum treated with fungi was not significantly altered; triacylglycerol was not detected in the cuticle extracts of any other tick species. Finally, A. sculptum and D. nitens cuticles have lipid components that may limit the development of M. robertsii.

Serosurvey of spotted fever group Rickettsia in equids from western Pará, Amazon, Brazil.

From a previous large epidemiological survey, we randomly selected 474 serum samples (463 horses and 11 mules) distributed among four municipalities of Pará state, Amazon region, Brazil, and from three types: farm animal, urban carthorse, and sport horse. Samples were tested by indirect fluorescence antibody test (IFAT ≥ 64) for antibodies reactive to spotted fever group (SFG) rickettsiae using Rickettsia rickettsii as crude antigens. From the 474 equids tested, 149 (31.4%) had ticks attached during sampling, belonging mostly to the species Dermacentor nitens. The overall seroprevalence for SFG rickettsiae was 31.4% (95% confidence interval: 27.3-35.9%) with 149 seropositive animals out of 474 screened. Notably, 77 equids (16.2%) had high endpoint titers ranging from 512 to 16,384, indicating that they had been exposed to SFG rickettsiae not long before sampling. Animal type affected rickettsial seroprevalence, with significantly higher values among farm horses when compared with urban and sport animals. Presence of dogs and tick infestation were negatively associated with equid seropositivity to R. rickettsii. This is the first report of SFG rickettsiae-reactive antibodies in equids from Pará state, Brazilian Amazon.

First molecular detection of Mycoplasma ovis in horses from Brazil

This study aimed to determine the occurrence of hemoplasmas and tick-borne pathogens (TBP) (Theileria equi, Babesia caballi, and Ehrlichia sp.) in horses and ticks’ salivary glands, and determine the factors associated with exposure/infection in a rural settlement in southern Brazil. Blood samples from 22 horses were screened for anti-T. equi and anti-Ehrlichia sp. antibodies by an indirect fluorescent antibody test (IFAT) assays. Samples were also tested by PCR assays for T. equi and B. caballi (18S rRNA and rap-1 genes, respectively), hemoplasmas (16S rRNA gene), and Ehrlichia sp. (dsb gene). Ticks were removed from the animals (inspection) and the environment (flannel trawling and dry ice traps), and morphologically identified. Additionally, salivary glands DNA was extracted from 28 adult ticks infesting the animals and four nymphs from the environment, and further screened for Ehrlichia sp. and hemotropic Mycoplasma sp. Anti-T. equi and anti-Ehrlichia sp. antibodies were detected in 40.91% (nine/22; 95% CI: 23.26-61.27) and 31.81% (seven/22; 95% CI: 16.36-52.68) horses, respectively. Theileria equi, B. caballi, and hemotropic Mycoplasma sp. DNA was detected in 59.09% (13/22), 4.55% (one/22), and 50% (11/22) horses, respectively. All horses tested negative in the PCR for Ehrlichia sp. All sequences showed ≥99% identity with multiple T. equi, B. caballi, and Mycoplasma ovis sequences deposited in GenBank database. Adult ticks were identified as Dermacentor nitens (44/47; 93.62%) and Rhipicephalus microplus (three/47; 6.38%). Ticks’ salivary glands were negative for Ehrlichia sp., while 39.29% from adults (11/28) and 50% from nymphs (two/four) from the environment were positive for hemotropic Mycoplasma sp. This is the first report of M. ovis infection in horses from Brazil and the first detection of hemoplasma DNA in salivary glands of D. nitens and R. microplus ticks. Further studies are needed to elucidate the vector competence of ticks to transmit hemoplasmas.

Ticks (Acari: Ixodidae) and Associated Pathoge Collected From Domestic Animals and Vegetation in Stann Creek District, Southeastern Belize, Central America.

Data on the prevalence and distribution of ticks and tick-borne diseases in Belize are lacking. Ticks (n = 564) collected from dogs, horses, and vegetation in two villages in Stann Creek District in southeastern Belize in 2018, were molecularly identified and screened for tick-borne nonviral human pathogens. The identity of 417 ticks was molecularly confirmed by DNA barcoding as Rhipicephalus sanguineus (Latreille) (66.43%), Amblyomma ovale Koch (15.59%), Dermacentor nitens Neumann (11.51%), Amblyomma sp. ADB0528 (3.6%), and the remainder being small records (2.87%) of Amblyomma coelebs Neumann, Amblyomma imitator Kohls, Amblyomma tapirellum Dunn, Amblyomma auricularium Conil, and Amblyomma maculatum Koch. Individual tick extracts were screened for the presence of Rickettsia spp., Babesia spp., Babesia microti, Borrelia spp., Ehrlichia spp., and Anaplasma spp. using available conventional polymerase chain reaction (PCR) assays. Rickettsia parkeri strain Atlantic Rainforest was identified in five specimens of A. ovale, and one other unidentified tick, all collected from dogs. Another unidentified tick—also collected from a dog—tested positive for an undefined but previously detected Ehrlichia sp. With the exception of D. nitens, all eight other tick species identified in this study were collected on dogs, suggesting that dogs could be usefully employed as sentinel animals for tick surveillance in Belize.

Molecular investigation of Babesia caballi in horses from the state of Rio de Janeiro, Brazil: Epidemiological aspects associated with the infection.

The epidemiological aspects of Babesia caballi infection were evaluated in 516 horse samples from Rio de Janeiro, Brazil. The presence and infestation level of ticks on horses, breed conditions, and animal management were evaluated on each farm through an epidemiological questionnaire. The gene that codes for rhoptry-associated protein-1 (RAP-1) of B. caballi was amplified by nested PCR (nPCR). Among the horses sampled, 17.2% (n=89/516) presented B. caballi DNA. The characterized samples showed 99-100% similarity with other isolates of B. caballi based on the RAP-1 gene, available in GenBank. In the final logistic regression model, the variables associated with B. caballi infection in horses were as follows: age below two years (OR=3.33; IC=1.7-6.5), farms located in low altitudes (OR=3.52; IC=1.7-7.3) and Dermacentor nitens infestation (OR=1.91; IC=1.1-3.4). Furthermore, a high level of D. nitens infestation in horses was also a factor associated with positivity for B. caballi (OR=2.11; IC=1.25-3.54). In summary, young horses bred in low altitude regions characterized with high temperatures, and infested by D. nitens, mainly with a higher level of infestation, are more likely to be infected by B. caballi. This epidemiological study provides statical evidence that the D. nitens tick play a role as the biological vector of B. caballi in the studied region.

Hard tick species (Acari: Ixodidae) and infestation in two livestock agroecosystems from Antioquia, Colombia.

Tick infestation affects about 80% of livestock globally while transmitting various pathogens causing high economic losses. This study aimed to determine the degree of tick infestation in two regions, North and Middle Magdalena in Antioquia, Colombia, to identify the ixodid tick species found and the associated risk factors. A cross-sectional study was carried out in 48 farms distributed in six municipalities of Antioquia. Two paddocks and eight bovines per farm were evaluated to estimate tick infestation (adults, nymphs, and larvae). Tick species were identified through a morphological and molecular analysis based on partial sequences of data obtained from DNA molecular markers, two mitochondrial (16S rRNA and COI), and one genomic DNA gene (18S rRNA). A multivariate Poisson regression model was applied to estimate the associated risk factors with ticks in cattle. Rhipicephalus microplus, Amblyomma patinoi and Dermacentor nitens were present in the livestock agroecosystems in the Middle Magdalena region; the highest incidence of tick infestation in cows and paddocks was reported in the municipality of Puerto Triunfo. The livestock agroecosystems in Middle Magdalena were characterized by a higher presence of adult R. microplus in cattle. Larval infestation of R. microplus, followed by D. nitens, was also found in paddocks. The multivariate analysis showed that the origin of cattle was the main risk factor associated with the presence of ticks (i.e., when cattle came from outside the farm). Cattle movement between farms in Middle Magdalena can contribute to the spread of ticks in this region.

First molecular detection of Spiroplasma spp. in ticks from horses in Brazil

The class Mollicutes comprises microorganisms that lack a cell wall, highly dependent on their host to survive. Within Mollicutes, the genus Spiroplasma comprises motile helical microorganisms associated with various insects and other arthropods. This study aimed to detect and characterize Mollicutes microorganisms in ticks of different species of veterinary importance, using molecular techniques. These ticks were collected from dogs, cats, cattle, and horses from Rio de Janeiro's metropolitan regions. They were morphologically classified and pooled according to their species for subsequent DNA extraction. These samples were tested by PCR using class Mollicutes-specific primers (16S rRNA) and positive amplicons were sequenced. The obtained DNA sequences were compared with other Mollicutes sequences deposited in GenBank. We found that four out of 745 (0.54%) of the tick pools were positive for members of the class Mollicutes, identified as Spiroplasma spp.; of the positive pools, one comprised Amblyomma sculptum adults and three comprised Dermacentor nitens nymphs. The present study describes Spiroplasma spp. in ticks in Brazil for the first time. Nevertheless, due to few reports on these microorganisms, further studies on epidemiology, virulence, and pathogenicity are needed.